A simple genotyping method for RIM15 gene polymorphisms in sake yeast strains
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Takashi Kuribayashi
Niigata Prefectural Sake Research Institute / Faculty of Food Industry, Niigata Agro-Food University
https://researchmap.jp/takashi-kuribayashi
- Nanako Asahi Faculty of Food Industry, Niigata Agro-Food University
- Hina Satone Faculty of Food Industry, Niigata Agro-Food University
- Jumpei Tanaka Niigata Prefectural Sake Research Institute
- Masamichi Sugawara Niigata Prefectural Sake Research Institute
- Keigo Sato Niigata Prefectural Sake Research Institute
- Yoshihito Nabekura Niigata Prefectural Sake Research Institute
- Toshio Joh Faculty of Agriculture, Niigata University
- Toshio Aoki Niigata Prefectural Sake Research Institute
DOI:
https://doi.org/10.51094/jxiv.868Keywords:
polymerase chain reaction-restriction fragment length polymorphism, RIM15, Saccharomyces cerevisiaeAbstract
In the process of brewing sake, Saccharomyces cerevisiae not only synthesizes numerous flavor compounds but also produces alcohol, which enhances fermentation efficiency. A previous study identified a sake yeast mutant with a single-nucleotide insertion in the RIM15 protein kinase gene (RIM15 with an insertion of adenine at position 5067: RIM15ins5067A). In this study, we developed primer sets (P1/P2) with engineered mismatches that introduce restriction sites for the restriction enzyme CpsCI. This allowed us to create a simple and rapid polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay to identify Kyokai no. 7 (K7) group sake yeast strains carrying the RIM15ins5067A mutation. We validated the PCR-RFLP method in detecting the RIM15ins5067A mutation in the K7 group of yeast strains. This approach provides a straightforward, effective, and valuable tool for distinguishing specific yeasts used in sake production.
Conflicts of Interest Disclosure
The authors declare no conflict of interest. All the experiments undertaken in this study complied with the current laws of the country in which they were performed.Downloads *Displays the aggregated results up to the previous day.
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Submitted: 2024-08-29 12:24:45 UTC
Published: 2024-09-03 02:22:20 UTC
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Copyright (c) 2024
Takashi Kuribayashi
Nanako Asahi
Hina Satone
Jumpei Tanaka
Masamichi Sugawara
Keigo Sato
Yoshihito Nabekura
Toshio Joh
Toshio Aoki
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